PLASMID ISOLATION AND ANALYSIS Part III Plasmid Isolation
- Slides: 11
PLASMID ISOLATION AND ANALYSIS Part III Plasmid Isolation
Plasmid Isolation Step 1 Place a QIAprep® spin column in the 2 ml collection tube. QIAprep® spin column collection tube PLASMID ISOLATION AND ANALYSIS: Part III
Plasmid Isolation Step 2 Use the small needle pipette to carefully remove all of the clear lysate/supernate from the “L” tube. PLASMID ISOLATION AND ANALYSIS: Part III
Plasmid Isolation Step 3 Add the clear lysate/supernate from tube “L” to the QIAprep® column. PLASMID ISOLATION AND ANALYSIS: Part III
Plasmid Isolation Step 4 Centrifuge for 1 minute and discard the flow through. PLASMID ISOLATION AND ANALYSIS: Part III
Plasmid Isolation Step 5 Using a small needle pipette, add the contents of the tube marked “Wash Buffer” (PE) to the QIAprep® spin column and centrifuge for 1 minute. PLASMID ISOLATION AND ANALYSIS: Part III
Plasmid Isolation Step 6 Discard the flow through, and centrifuge for an additional 1 minute to remove residual wash buffer. PLASMID ISOLATION AND ANALYSIS: Part III
Plasmid Isolation Step 7 Transfer the QIAprep® spin column to the 1. 5 ml tube marked “M” in Part II, step 6. QIAprep® spin column PLASMID ISOLATION AND ANALYSIS: Part III
Plasmid Isolation Step 8 Use a small needle pipette to elude the DNA by adding the contents of the tube marked sterile water (SW) to the QIAprep® spin column. PLASMID ISOLATION AND ANALYSIS: Part III
Plasmid Isolation Step 9 Let stand for 1 minute and centrifuge for 1 minute to collect the DNA. PLASMID ISOLATION AND ANALYSIS: Part III
Plasmid Isolation Step 10 Label the “M” tube with your group’s name, seal the tube with its original cap, and place in freezer (-20 o. C) until you are ready to analyze the extracted DNA in Part IV. PLASMID ISOLATION AND ANALYSIS: Part III